作者感谢Laura Tucker和Natasa Loncarevic对手稿的评论以及Instituto de Medicina Molecular João Lobo Antunes的啮齿动物设施给予的支持。作者希望感谢Prémios Santa Casa Neurociências - 脊髓损伤研究奖Melo e Castro（MC-36/2020）对L.S.和C.S.M.的财政支持。这项工作得到了科学与技术基金会（FCT）（PTDC/BIA-COM/0151/2020）、iNOVA4Health（UIDB/04462/2020和UIDP/04462/2020）和LS4FUTURE（LA/P/0087/2020）的支持，得到了CEEC个人首席研究员合同（2021.02253.CEECIND）的支持。AFI得到了FCT（2020.08168.BD）的博士奖学金的支持。AMM得到了FCT博士奖学金的支持（PD/BD/128445/2017）。I.M.得到了FCT的博士后奖学金（SFRH / BPD / 118051 / 2016）的支持。DNS得到了FCT博士奖学金的支持（SFRH / BD / 138636 / 2018）。

所有处理、手术和术后护理程序均由医学研究所分子内部委员会 （ORBEA） 和葡萄牙动物伦理委员会 （DGAV） 根据欧洲共同体指南（指令 2010/63/EU）和葡萄牙动物护理法 （DL 113/2013） 批准，许可证为 0421/000/000/2022。本研究使用9周龄的雌性C57Bl / 6J小鼠。尽一切努力尽量减少动物的数量并减少研究中使用的动物的痛苦。MATLAB脚本和MW软件的独立版本是开源的，可在GitHub上获得。 存储库 （<a href="https://github.com/NeurogeneLocomotion/MouseWalker">https://github.com/NeurogeneLocomotion/MouseWalker</a>）。虽然MW软件是在MATLAB R2012b中开发的，但它已经适应在MATLAB R2022b中运行。 图1 显示了MW的分析工作流程。
1. 设置鼠标步行者 （MW） 设备
<ol><li>如前所述组装MW装置23，或适应实验设计的特定需求（有关设置的更多详细信息，请参见 材料表 和 补充图1 ）。 注意：步行竞技场可以做得更宽，以容纳更大的动物，如老鼠。</li>
	<li>
	<div dir="auto" style="color: rgb(34, 34, 34); font-family: Arial, Helvetica, sans-serif; font-size: small; background-color: rgb(255, 255, 255);">验证动物行走的有机玻璃是否干净且无划痕。使用光滑的清洁布，并尽量减少使用高浓度的有机溶剂，如氨或乙醇，这可能会损坏有机玻璃（建议使用3%过氧化氢，7%乙醇或任何兼容且适当的有机玻璃消毒剂）。如有必要，请更换有机玻璃。</div>
	</li>
	<li>为高速摄像机设置快速镜头和大光圈（即较小的光圈值）以捕获大量光线，因为这有助于记录fTIR信号（参见 材料表）。 注意：镜头不应产生光学失真，尤其是在图像边缘。可以通过记录已知图案（例如条纹或正方形）然后在 ImageJ/FIJI 24 上测量块的大小来测试光学畸变（使用线条工具，然后单击分析 >测量）。例如，一个 1 厘米大小的正方形在图像中心和边缘应具有相同的像素尺寸。变化应小于 5%。</li>
	<li>从背景灯箱点亮五彩 LED 灯条。</li>
	<li>点亮人行道灯箱上的白色 LED 灯条。 注意：彩色LED也可以使用25 ，以方便区分封装/主体/背景。</li>
	<li>在房间灯熄灭的情况下，验证背景灯箱和走道的光强度。如有必要，使用电位计或半透明塑料调整强度。这些必须进行优化，以便像素强度按以下顺序增加：动物的身体<背景<脚印。<ol><li>要检查动物身体/背景/脚印的像素强度，请在 ImageJ/FIJI24 上打开图像序列，然后单击分析 >测量。封装信号不应过饱和，因为这会阻止定义封装的边界（即脚趾和脚垫）（补充图2）。</li>
	</ol></li>
	<li>在视频录制软件上调整人行道的图像对比度。对比度可以通过两种方式进行调整：调暗或增加 LED 灯条上的照明，或者调整相机镜头光圈。</li>
	<li>正确放置镜头，使其处于相同的高度，位于 45° 反射镜的中心，并与人行道垂直 （90°）。这将沿左右走道生成一个恒定比例的图像。 注意：避免在多个录制会话中更改摄像机位置（距离、高度和方向）。如有必要，标记应放置三脚架的地板。这将保持图像功能。</li>
	<li>将镜头聚焦在有机玻璃表面。这可以使用接触有机玻璃表面的非破坏性物体进行测试。 注意：使用较低的光圈镜头值，景深会变小，从而使对焦更加困难。</li>
	<li>确保在测定过程中所有设置保持不变，因为它们可能会改变录制视频的像素强度。</li>
</ol>2. 视频采集
<ol><li>在测试之前，确保小鼠熟悉房间和设备。至少保存 1 天用于习惯化（第 0 天）。为避免过度训练，请在与其他行为测试不同的日期（最好是第二天）进行MW测试。</li>
	<li>在视频录制软件中，确保至少 50 厘米的人行道可见。</li>
	<li>调整录制设置以截断人行道区域。这将减小视频大小并优化视频采集。</li>
	<li>在每次会议前拍摄常规尺子的照片或短视频。稍后将在“设置窗口”中使用每厘米的像素数来校准视频。</li>
	<li>开始视频采集，抓住尾巴根部将动物放在走道边缘，以免受伤。确保动物向前移动到平台的最边缘。以至少 100 帧/秒的速度执行视频录制，以确保平稳的步态过渡。<ol><li>如果需要，通过轻轻敲击走道墙壁或打响/拍打手指来激励动物移动。但是，请避免物理轻推，因为这可能会影响结果。</li>
		<li>将视频直接保存为 TIFF （使用 LZW 压缩）、JPEG 或 PNG 格式的图像序列。如果摄像机录制为原始MOV文件，请在ImageJ / FIJI24 中打开文件并单击 文件>另存为 图像序列（或使用其他软件，例如LosslessCut25），将视频转换为图像序列>。 注意：大多数动物在被放入人行道后立即开始行走;因此，建议在放置动物之前开始视频采集。</li>
	</ol></li>
</ol>3. 为 MW 跟踪软件准备视频
<ol><li>拍摄足够多的每只鼠标的完整运行。每个条件要拍摄的动物数量和完整运行的数量必须根据每个实验设计决定。完全运行是指鼠标在没有长时间停止的情况下走过整个 50 厘米的人行道（在本实验中，选择了三个完整的运行）。 注意：根据图像采集软件的不同，可能需要将视频裁剪到最小的ROI。这将提高跟踪和输出生成的速度。</li>
	<li>在 ImageJ/FIJI24 中，通过单击图像>“堆栈” >“工具”>“创建子堆栈”，选择鼠标在屏幕上的帧。MW上的跟踪要求头部和尾部在所有帧中都可见。但是，可以从单个视频录制中创建多个子堆栈，这些子堆栈稍后将表示每次运行。</li>
	<li>通过单击“文件”>“另 存为图像序列”，将每个子堆栈分别保存在不同的文件夹中>。MW 软件随后会在每次开始分析运行时自动在每个目录中创建一个子文件夹。</li>
</ol>4. 追踪
<ol><li>打开 MATLAB，将包含 MW 脚本的文件夹添加到工作目录，然后在主命令行上运行 “MouseWalker.m”。 注意：使用 MATLAB 下的 MW 软件可以在 MATLAB 的主控制台上查看跟踪错误消息并选择所需的输出数据（通过打开主脚本文件 “MouseEvaluate.m” 并将输出更改为 1 或 0：excel 文件、脚步图、姿势轨迹和步态模式）。</li>
	<li>将视频文件夹加载为“输入目录”。还可以选择输出文件夹;但是，这不是必需的，因为MW软件会在“输入目录”中自动创建一个名为“结果”的新文件夹。</li>
	<li>使用箭头“<<”、“<”、“>>”和“>”检查视频帧是否全部正确加载到 MW 软件中。</li>
	<li>转到所有校准和阈值参数所在的“设置窗口”。这些设置可能会根据背景和足迹的像素强度以及主体和足迹的最小尺寸以及其他因素而变化（请参阅 补充图 2 中的示例）。通过单击“ 预览 ”按钮来测试更改某些参数的效果。<ol><li>使用不同的绘图样式，包括“身体+脚+尾巴”、“仅身体”、“仅脚”和“仅尾巴”，以帮助区分调整阈值参数后的身体部位。</li>
		<li>利用右侧面板上的工具测量亮度或大小（分别使用“亮度”和“标尺”按钮）。只要相机距离保持不变，所有设置都可以保存为“默认”。</li>
	</ol></li>
	<li>调整阈值参数后，检查视频是否已准备好进行自动跟踪。转到第一帧，然后单击 自动 开始跟踪。可以实时执行此步骤，并且需要几分钟，具体取决于视频的大小和计算机的性能。<ol><li>如果自动跟踪错误地标记了身体特征，请取消自动跟踪，输入新设置，然后重新启动该过程。</li>
	</ol></li>
	<li>跟踪完成后，检查是否需要手动更正。要更正，请使用中间面板选择或取消选择，并指示右前 （RF）、右后部 （RH）、左前 （LF） 和左后后 （LH） 爪脚印、头部、鼻子、身体（分为两段）和尾部位置（分为四段）的位置。按“保存”按钮 保存 更改。 注意：所有按钮和大多数命令都有一个快捷键（有关详细信息，请查看相关手册23）。为了方便视频滚动和键盘快捷键的执行，可以使用带有可编程按钮和穿梭轮的硬件控制器，如 Contour ShuttlePro V2。</li>
	<li>点击 评估 从跟踪的视频生成输出文件。根据所选的所需输出（请参阅步骤 4.1），此步骤可能需要几分钟时间。</li>
	<li>检查所有图形输出数据图是否保存在“结果”文件夹中。通过检查一些图形输出来验证跟踪的准确性，例如“立场轨迹”，其中可以检查所有爪子位置是否一致。<ol><li>如果发现错误，请手动更正跟踪（如果可能;否则，请删除“结果”文件夹，然后使用新设置再次执行自动跟踪），然后再次单击 “评估 ”命令。</li>
	</ol></li>
	<li>检查MW软件生成的所有定量测量值是否保存在Excel电子表格中，并汇总在“1.Info_Sheet”。确保公式分隔的 excel 选项与脚本匹配。小数点分隔符必须是“，”，千位分隔符必须是“;”。</li>
	<li>使用“MouseMultiEvaluate.m”脚本将所有运行的测量值聚集到一个新文件中进行分析。<ol><li>首先，生成一个包含所有视频的文件夹路径的.txt文件（例如“视频文件.txt”）。确保每行对应于一个视频。</li>
		<li>然后，将“MouseMultiEvaluate（'Videofiles.txt'）”写入命令行。将在工作目录中生成一个名为“ResultSummary.xls”的 excel 文件（请参阅 GitHub 存储库中的示例）。 注意： 图2 表示MW软件从一只记录的动物的视频中获得的图形输出。</li>
	</ol></li>
</ol>5. 运动学数据分析工作流程
<ol><li>根据以下先决条件，编辑步骤 4.10 中生成的 Excel 工作表，其中包含要使用提供的 Python 脚本进行处理的数据。<ol><li>在第一列标题中，指定实验条件。为组/条件名称后面的每一行命名（来自同一组的个人必须具有相同的名称）。第一组必须是控件或基线（这仅对于热图绘图（步骤 5.6）是必需的）。</li>
		<li>在第二列中，指定动物 ID。这是强制性的，尽管此信息不会用于绘图生成。</li>
		<li>在第三列以后，选择将用于分析的电机参数。确保第一行是参数的名称（这些名称稍后将出现在图中）。</li>
	</ol></li>
	<li>打开 Anaconda Navigator，然后执行 Spyder 以打开提供的 Python 脚本。 注意：所有脚本都是用Python 3.9.13开发的，在Anaconda Navigator 2.1.4中使用Spyder 5.2.2执行，并且可以在材料表和GitHub存储库中找到（其中包括其他 材料 ，例如视频示例，Excel示例文件和FAQ文档）。可以在 Anaconda Navigator 之外执行脚本;但是，此图形用户界面更加用户友好。</li>
	<li>使用“Rawdata_PlotGenerator.py”生成原始数据图。这将允许将每个参数可视化为速度的函数。<ol><li>在Spyder中打开“Rawdata_PlotGenerator.py”，然后单击 “播放 ”按钮运行代码。</li>
		<li>在自动窗口中选择要分析的 Excel 文件和工作表名称。如果工作表名称未更改，请填写“Sheet1”。</li>
		<li>原始数据图将显示在绘图控制台（右上方面板）中。要保存绘图，请单击绘图控制台中的 保存 图像或 保存所有图像 按钮。</li>
	</ol></li>
	<li>使用脚本“Residuals_DataAnalysis”计算用于数据分析的残差。此脚本将生成一个 CSV 文件，其中包含所有电机参数的残差计算。 注意：MW 提取的许多测量步态参数随速度而变化（例如，摆动速度、步长、站立持续时间、站姿直线度和步态指数）。因此，建议对基线实验的每个参数与速度执行最佳拟合回归模型，然后确定与此回归模型相关的每个实验组的残差值。然后将数据表示为与残差归一化线26 的差值。<ol><li>在Spyder中打开“Residuals_DataAnalysis.py”，然后单击 “播放 ”按钮运行代码。</li>
		<li>在自动窗口中选择要分析的 Excel 文件和工作表名称。如果工作表名称未更改，请填写“Sheet1”。</li>
		<li>将 CSV 文件保存在与数据相同的文件夹中。控件（或基线）必须是 Excel 文件中的第一个组。</li>
	</ol></li>
	<li>使用“PCA_PlotGenerator.py”脚本执行主成分分析 （PCA）。 注意：这种无监督降维方法用于生成更简洁的数据表示27，28，29（图3A，B）。PCA 脚本包括以下步骤。首先通过居中和缩放对数据进行预处理，然后PCA算法计算协方差矩阵以确定变量之间的相关性，并计算协方差矩阵的特征向量和特征值以识别主成分。选择前两个或三个主分量分别用于在 2D 或 3D 图中表示数据。图中的每个点对应于一种动物，并代表一个不同的抽象变量。颜色编码的点用于区分特定组。因此，点簇反映了相应个体共享的相似行走模式。<ol><li>在Spyder中打开“PCA_PlotGenerator.py”，然后单击 “播放 ”按钮运行代码。</li>
		<li>在自动窗口中选择要分析的 Excel 文件和工作表名称。如果工作表名称未更改，请填写“Sheet1”。</li>
		<li>确保 PCA 2D 和 3D 图显示在打印控制台（右上方面板）中。每种颜色代表不同的组，图例显示在绘图旁边。要保存绘图，请单击绘图控制台中的 保存图像 。</li>
	</ol></li>
	<li>使用“Heatmap_PlotGenerator.py”生成热图。确保热图生成器创建一个表，显示每个电机参数27 的基线组（或对照组）与其他组之间的统计差异（图4）。每列描绘一个组，每行与特定的电机参数相关。 注意：统计分析是使用单因素方差分析进行的，然后是Tukey的事后检验（用于正态分布）或Kruskal-Wallis 方差分析，然后是邓恩事后检验（对于非正态分布）。异常值被排除在分析之外。 P 值由颜色代码表示，红色和蓝色阴影分别表示相对于对照（或基线）的增加或减少。色阴影表示统计显著性，较深的颜色表示较高的显著性，较浅的颜色表示较低的显著性。对应于 P < 0.001;** 对应于 P < 0.01;和 * 对应于 P < 0.05。白色表示无变化。<ol><li>在Spyder中打开“Heatmap_PlotGenerator.py”，然后单击 “播放 ”按钮运行代码。</li>
		<li>在自动窗口中选择要分析的 Excel 文件和工作表名称。如果工作表名称未更改，请填写“Sheet1”。</li>
		<li>在第二个自动窗口中选择数据类型：原始数据或残差数据。如果未选择某个选项，则残差数据为默认值。</li>
		<li>热图将显示在绘图控制台（右上方面板）中。要保存绘图，请单击绘图控制台中的 保存图像 。 注意：控件（或基线）必须是 Excel 文件中的第一个组。</li>
	</ol></li>
	<li>使用“Boxplots_PlotGenerator.py”生成箱线图。该工具将允许生成箱线图，表示每组所有电机参数的值分布（图 5、图 6 和图 7）。 注意：每个框都包含中位数作为中间线，框的下边缘和上边缘分别表示 25% 和 75% 的四分位数。晶须表示完整数据集的范围，不包括异常值。异常值定义为分别低于或高于 25% 和 75% 四分位数的四分位数范围 1.5 倍的任何值。<ol><li>在Spyder中打开“Boxplots_PlotGenerator.py”，然后单击 “播放 ”按钮运行代码。</li>
		<li>在自动窗口中选择要分析的 Excel 文件和工作表名称。如果工作表名称未更改，请填写“Sheet1”。</li>
		<li>在第二个自动窗口中选择数据类型：原始数据或残差数据。如果未选择某个选项，则残差数据为默认值。</li>
		<li>箱线图将显示在绘图控制台（右上方面板）中。要保存绘图，请单击绘图控制台中的 保存 图像或 保存所有图像 按钮。</li>
	</ol></li>
</ol>

使用 MouseWalker 量化脊髓损伤小鼠的运动功能障碍

<ol>
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提交人声明，他们没有相互竞争的经济利益。

在这里，通过分析SCI后的运动行为来证明MouseWalker方法的潜力。它为步进、足迹和步态模式的特定变化提供了新的见解，否则其他标准测试会错过这些变化。除了提供 MW 包的更新版本外，还使用提供的 Python 脚本描述了数据分析工具（请参阅步骤 5）。
由于MW生成了一个大型数据集和反映高维运动过程的运动学参数集合，因此采用了PCA;事实上，PCA已被广泛用于其他运动学数据?...

四肢的有效协调并非四足动物所独有。人类的前肢-后肢协调对于完成多项任务仍然很重要，例如游泳和行走时的速度改变1.各种肢体运动学2 和运动程序1，3，4 以及本体感觉反馈回路5 在人类和其他哺乳动物之间是保守的，在分析运动障碍（例如脊髓损伤 （SCI）6，7，8）的治疗方案时应考虑这些选择。
为了行走，来自前肢和后肢的几个脊柱连接需要正确连接和有节奏地激活，这需要来自大脑的输入和来自体感系统的反馈2，9，10。这些连接在中央模式发生器（CPG）中达到顶峰，它们分别位于前肢和后肢的颈椎和腰椎水平1，9，10。通常，在 SCI 之后，神经元连接的破坏和抑制性神经胶质瘢痕12 的形成限制了运动功能的恢复，结果从完全瘫痪到一组肢体功能受限不等，具体取决于损伤的严重程度。精确量化 SCI 后运动功能的工具对于监测恢复和评估治疗或其他临床干预措施的效果至关重要6.
SCI 小鼠挫伤模型的标准度量测定是巴索小鼠量表 （BMS）13，14，这是一种非参数评分，考虑了空旷场地的躯干稳定性、尾部位置、足底步和前肢-后肢协调。尽管BMS在大多数情况下非常可靠，但它需要至少两名经验丰富的评分员来观察动物运动的所有角度，以解释自然变异性并减少偏差。
还开发了其他测定方法来定量评估SCI后的运动性能。其中包括旋转测试，它测量在旋转圆柱体上花费的时间15;水平梯子，测量错过栏杆和正梯子抓斗的数量16，17;以及横梁行走测试，该测试测量动物在穿越窄梁时所花费的时间和失败次数18.尽管反映了运动缺陷的组合，但这些测试都没有产生关于前肢 - 后肢协调的直接运动信息。
为了具体和更彻底地分析步行行为，已经开发了其他测定方法来重建步数循环和步态策略。一个例子是足迹测试，其中动物的墨水爪子在一张白纸上画一个图案19.虽然执行简单，但提取步幅等运动学参数既麻烦又不准确。此外，缺乏动态参数，例如步进周期的持续时间或腿定时协调，限制了其应用;事实上，这些动态参数只能通过分析啮齿动物穿过透明表面的逐帧视频来获取。对于SCI研究，研究人员使用跑步机从侧面分析了步行行为，包括重建步数循环和测量每个腿关节的角度变化4，20，21。尽管这种方法可以提供非常丰富的信息6，但它仍然专注于一组特定的肢体，并且缺乏额外的步态特征，例如协调性。
为了填补这些空白，Hamers及其同事开发了一种基于光学触摸传感器的定量测试，使用受挫全内反射（fTIR）22。在这种方法中，光线通过内部反射 穿过 玻璃传播，在按压爪子时散射，最后被高速相机捕获。最近，这种方法的开源版本，称为MouseWalker，已经可用，这种方法将fTIR走道与跟踪和量化软件包23相结合。使用这种方法，用户可以提取大量定量参数，包括步数、空间和步态模式、脚印定位和前肢-后肢协调，以及视觉输出，例如脚印模式（模仿墨水爪测定6）或相对于身体轴的站立阶段。重要的是，由于其开源性质，可以通过更新 MATLAB 脚本包来提取新参数。
在这里，更新了以前发布的 MouseWalker23 系统组件。提供了如何设置它的说明，以及实现最佳视频质量、跟踪条件和参数采集所需的所有步骤。还共享了其他量化后工具，以增强对MouseWalker（MW）输出数据集的分析。最后，通过在脊髓损伤（SCI）背景下获得一般运动性能的可量化值，特别是步道周期和前肢 - 后肢协调，证明了该工具的有用性。

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			<td>45&#186; Mirror&#160;</td>
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			<td></td>
			<td></td>
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			<td>2 aluminum extrusion (2 x 2 cm), 16 cm height, 1 on each side</td>
			<td>Misumi</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2 aluminum extrusion (2 x 2 cm), 23 cm, @ 45&#176; , 1 on each side</td>
			<td>Misumi</td>
			<td></td>
			<td></td>
		</tr>
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			<td>1 aluminum extrusion (2 x 2 cm), 83 cm long</td>
			<td>Misumi</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>87 x 23 cm mirror</td>
			<td>General glass supplier&#160;</td>
			<td></td>
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		</tr>
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			<td>black cardboard filler&#160;</td>
			<td>General stationery supplier</td>
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			<td>We used 2, one with 69 x 6 cm and another with 69 x 3cm to limit the reflection on the mirror</td>
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			<td>Background backlight</td>
			<td></td>
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			<td>109 x 23 cm plexiglass (0.9525 cm thick)</td>
			<td>General hardware supplier</td>
			<td></td>
			<td></td>
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			<td>2 lateral aluminum extrusion (4 x 4 cm), 20 cm long, 1 on each side</td>
			<td>Misumi</td>
			<td></td>
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			<td>multicolor LED strip</td>
			<td>General hardware supplier</td>
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			<td>white opaque paper to cover the plexyglass</td>
			<td>General stationery supplier</td>
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			<td>fTIR Support base and posts</td>
			<td></td>
			<td></td>
			<td></td>
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			<td>2 aluminum extrusion (4 x 4 cm), 100 cm height</td>
			<td>Misumi</td>
			<td></td>
			<td></td>
		</tr>
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			<td>60 x 30 cm metric breadboard</td>
			<td>Edmund Optics&#160;</td>
			<td>#54-641</td>
			<td></td>
		</tr>
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			<td>M6 12 mm screws</td>
			<td>Edmund Optics&#160;</td>
			<td></td>
			<td></td>
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			<td>M6 hex nuts and wahers</td>
			<td>Edmund Optics&#160;</td>
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			<td>fTIR Walkway&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
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			<td>109 x 8.5 cm plexyglass (1.2 cm thick)</td>
			<td>General hardware supplier</td>
			<td></td>
			<td>109 x 8.5 cm plexyglass (1.2 cm thick)</td>
		</tr>
		<tr>
			<td>109 cm long Base-U-channel aluminum with 1.6 cm height x 1.9 cm depth thick folds (to hold the plexyglass)</td>
			<td>General hardware supplier</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2 lateral aluminum extrusion (4 x 4 cm) 20 cm length, 1 on each side</td>
			<td>Misumi</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>black cardboard filler&#160;</td>
			<td>General stationery supplier</td>
			<td></td>
			<td>we used 2 fillers on each side to cover the limits of the plexyglass, avoiding bright edges</td>
		</tr>
		<tr>
			<td>12 mm screws</td>
			<td>Edmund Optics&#160;</td>
			<td>M6</td>
			<td></td>
		</tr>
		<tr>
			<td>High speed camera (on a tripod)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Blackfly S USB3</td>
			<td>Blackfly</td>
			<td>USB3</td>
			<td>This is a reccomendation. The requirement is to record at least 100 frames per second</td>
		</tr>
		<tr>
			<td>Infinite Horizon Impactor&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Infinite Horizon Impactor&#160;</td>
			<td>Precision Systems and Instrumentation, LLC.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Lens</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Nikkon AF Zoom-Nikkor 24-85mm</td>
			<td>Nikkon&#160;</td>
			<td>2.8-4D IF</td>
			<td>This lens is reccomended, however other lens can be used. Make sure it contains a large aperture (i.e., smaller F-stop values), to capture fTIR signals</td>
		</tr>
		<tr>
			<td>Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MATLAB R2022b</td>
			<td>MathWorks</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Python 3.9.13&#160;</td>
			<td>Python Software Foundation</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anaconda Navigator 2.1.4</td>
			<td>Anaconda, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spyder 5.1.5&#160;</td>
			<td>Spyder Project Contributors</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Walkway wall&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2 large rectagular acrilics with 100 x 15 cm</td>
			<td>Any bricolage convenience store</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2 Trapezian acrilic laterals with 6-10 length x 15 cm height</td>
			<td>Any bricolage convenience store</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>GitHub Materials</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Folder name</td>
			<td>URL</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Boxplots</td>
			<td>https://github.com/NeurogeneLocomotion/MouseWalker/tree/main/Boxplots</td>
			<td></td>
			<td>Script to create Boxplots</td>
		</tr>
		<tr>
			<td>Docs</td>
			<td>https://github.com/NeurogeneLocomotion/MouseWalker/tree/main/Docs</td>
			<td></td>
			<td>Additional documents</td>
		</tr>
		<tr>
			<td>Heatmap</td>
			<td>https://github.com/NeurogeneLocomotion/MouseWalker/tree/main/Heatmaps</td>
			<td></td>
			<td>Script to create heatmap</td>
		</tr>
		<tr>
			<td>Matlat script</td>
			<td>https://github.com/NeurogeneLocomotion/MouseWalker/tree/main/Matlab%20Script</td>
			<td></td>
			<td>MouseWalker matlab script</td>
		</tr>
		<tr>
			<td>PCA</td>
			<td>https://github.com/NeurogeneLocomotion/MouseWalker/tree/main/PCA%20plots</td>
			<td></td>
			<td>Script to perform Principal Component Analysis</td>
		</tr>
		<tr>
			<td>Raw data Plots</td>
			<td>https://github.com/NeurogeneLocomotion/MouseWalker/tree/main/Rawdata%20Plots</td>
			<td></td>
			<td>Script to create Raw data plots</td>
		</tr>
		<tr>
			<td>Residual Analysis</td>
			<td>https://github.com/NeurogeneLocomotion/MouseWalker/tree/main/Residual_Analysis</td>
			<td></td>
			<td>Code to compute residuals from Raw data</td>
		</tr>
	</tbody>
</table>

using the mousewalker to quantify locomotor dysfunction in a mouse model of spinal cord injury

复杂且高度协调的运动程序（例如步行和跑步）的执行取决于脊柱和脊髓上回路的节律激活。胸椎脊髓损伤后，与上游回路的通信受损。这反过来又导致协调的丧失，恢复潜力有限。因此，为了更好地评估给予药物或治疗后的恢复程度，有必要使用新的、更详细和准确的工具来量化脊髓损伤动物模型中步态、肢体协调和其他运动行为的精细方面。多年来，已经开发了几种测定方法来定量评估啮齿动物的自由行走行为;然而，它们通常缺乏与步进步态策略、足迹模式和协调相关的直接测量。为了解决这些缺点，提供了MouseWalker的更新版本，它将受挫的全内反射（fTIR）走道与跟踪和量化软件相结合。这个开源系统已经适应于提取几个图形输出和运动学参数，并且一组后量化工具可以分析提供的输出数据。这份手稿还展示了这种方法如何与已经建立的行为测试相结合，定量描述脊髓损伤后的运动缺陷。

标准BMS系统描述了SCI14之后的总运动缺陷。由于其主观性，其他定量测定通常与BMS一起进行，以产生更详细和精细的运动评估。然而，这些测试未能显示有关步步周期、步进模式和前肢-后肢协调的具体信息，这对于了解脊柱回路如何维持功能并适应不完全的 SCI 非常重要。本节展示了MW工具箱如何帮助监测SCI后运动功能的恢复，并添加有关步行行为的相关信息。
<p class="jove_cont...

Watch this Scientific Journal Video about Using the MouseWalker to Quantify Locomotor Dysfunction in a Mouse Model of Spinal Cord Injury at JoVE.com

作者聚焦：使用 MouseWalker 量化脊髓损伤小鼠模型中的运动功能障碍  

提供了一个实验管道，用于使用MouseWalker（MW）工具箱定量描述自由行走的小鼠的运动模式，范围从初始视频记录和跟踪到定量后分析。使用小鼠脊髓挫伤损伤模型来证明MW系统的有用性。

使用MouseWalker量化脊髓损伤小鼠模型中的运动功能障碍

The execution of complex and highly coordinated motor programs, such as walking and running, is dependent on the rhythmic activation of spinal and ...

在这项研究中，我们使用脊髓损伤的小鼠模型来揭示促进脊髓修复的方法，这些方法可能导致有效的运动和功能感觉恢复。使用鼠标步行器，我们可以通过将几个图形输出和运动学参数与一组后定量工具相结合，对脊髓恢复进行更定量的分析。高质量的摄像机和复杂的软件包可以更直接和定量地描述运动活动。鼠标步行者就是一个很好的例子。目前存在两个主要挑战。首先，实验室在其实验管道中实施这些技术的能力。其次，减少从视频录制到数据生成的时间和步骤。我们意识到标准方案不足以描述我们在脊髓损伤后看到的所有机车缺陷。我们还发现，鼠标步行器帮助我们测量协调性，这是标准测试中最难评估的参数之一。在这里，软件不仅是免费和开源的，而且硬件也很容易用简单的材料组装。我们还提供开源工具来分析鼠标步行者数据。鼠标步行器是研究其他运动相关或运动功能障碍的宝贵方法，而不仅仅是脊髓损伤。它也可以与已经建立的行为协议结合使用。

using-mousewalker-to-quantify-locomotor-dysfunction-mouse-model

Research

JoVE Journal

Neuroscience

Using the MouseWalker to Quantify Locomotor Dysfunction in a Mouse Model of Spinal Cord Injury

2.5K 次观看.  Faculdade de Medicina da Universidade de Lisboa. 在这项研究中，我们使用脊髓损伤的小鼠模型来揭示促进脊髓修复的方法，这些方法可能导致有效的运动和功能感觉恢复。使用鼠标步行器，我们可以通过将几个图形输出和运动学参数与一组后定量工具相结合，对脊髓恢复进行更定量的分析。高质量的摄像机和复杂的软件包可以更直接和定量地描述运动活动。鼠标步行者就是一个很好的例子。目前存在两个主要挑战。?...

视频: 作者聚焦：使用 MouseWalker 量化脊髓损伤小鼠模型中的运动功能障碍  

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has significantly broadened our knowledge of physiological and pathological processes in numerous tissues in vivo 4-7. In studies of the central nervous system (CNS), there has been a broad application of in vivo imaging in the brain, which has produced a plethora of novel and often unexpected findings about the behavior of cells such as neurons, astrocytes, microglia, under physiological or pathological conditions 8-17. However, mostly technical complications have limited the implementation of in vivo imaging in studies of the living mouse spinal cord. In particular, the anatomical proximity of the spinal cord to the lungs and heart generates significant movement artifact that makes imaging the living spinal cord a challenging task.
We developed a novel method that overcomes the inherent limitations of spinal cord imaging by stabilizing the spinal column, reducing respiratory-induced movements and thereby facilitating the use of two-photon microscopy to image the mouse spinal cord in vivo. This is achieved by combining a customized spinal stabilization device with a method of deep anesthesia, resulting in a significant reduction of respiratory-induced movements. This video protocol shows how to expose a small area of the living spinal cord that can be maintained under stable physiological conditions over extended periods of time by keeping tissue injury and bleeding to a minimum. Representative raw images acquired in vivo detail in high resolution the close relationship between microglia and the vasculature. A timelapse sequence shows the dynamic behavior of microglial processes in the living mouse spinal cord. Moreover, a continuous scan of the same z-frame demonstrates the outstanding stability that this method can achieve to generate stacks of images and/or timelapse movies that do not require image alignment post-acquisition. Finally, we show how this method can be used to revisit and reimage the same area of the spinal cord at later timepoints, allowing for longitudinal studies of ongoing physiological or pathological processes in vivo.

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

A minimally invasive protocol to stabilize the mouse spinal column and perform repetitive in vivo spinal cord imaging using two-photon microscopy is described. This method combines a spinal stabilization device and an anesthetic regimen to minimize respiratory-induced movements and produce raw imaging data that require no alignment or other post-processing.

在体内脊髓的鼠标线使用双光子显微镜成像

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has ...

科研

神经科学

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and mortality. Based on the nature of primary injury following TBI, complex and heterogeneous secondary consequences result, which are followed by regenerative processes 1,2. Primary injury can be induced by a direct contusion to the brain from skull fracture or from shearing and stretching of tissue causing displacement of brain due to movement 3,4. The resulting hematomas and lacerations cause a vascular response 3,5, and the morphological and functional damage of the white matter leads to diffuse axonal injury 6-8. Additional secondary changes commonly seen in the brain are edema and increased intracranial pressure 9. Following TBI there are microscopic alterations in biochemical and physiological pathways involving the release of excitotoxic neurotransmitters, immune mediators and oxygen radicals 10-12, which ultimately result in long-term neurological disabilities 13,14. Thus choosing appropriate animal models of TBI that present similar cellular and molecular events in human and rodent TBI is critical for studying the mechanisms underlying injury and repair.
Various experimental models of TBI have been developed to reproduce aspects of TBI observed in humans, among them three specific models are widely adapted for rodents: fluid percussion, cortical impact and weight drop/impact acceleration 1. The fluid percussion device produces an injury through a craniectomy by applying a brief fluid pressure pulse on to the intact dura. The pulse is created by a pendulum striking the piston of a reservoir of fluid. The percussion produces brief displacement and deformation of neural tissue 1,15. Conversely, cortical impact injury delivers mechanical energy to the intact dura via a rigid impactor under pneumatic pressure 16,17. The weight drop/impact model is characterized by the fall of a rod with a specific mass on the closed skull 18. Among the TBI models, LFP is the most established and commonly used model to evaluate mixed focal and diffuse brain injury 19. It is reproducible and is standardized to allow for the manipulation of injury parameters. LFP recapitulates injuries observed in humans, thus rendering it clinically relevant, and allows for exploration of novel therapeutics for clinical translation 20.
We describe the detailed protocol to perform LFP procedure in mice. The injury inflicted is mild to moderate, with brain regions such as cortex, hippocampus and corpus callosum being most vulnerable. Hippocampal and motor learning tasks are explored following LFP.

Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is described.

侧向液压冲击：创伤性脑损伤小鼠模型

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

An Ex Vivo Laser-induced Spinal Cord Injury Model to Assess Mechanisms of Axonal Degeneration in Real-time

Injured CNS axons fail to regenerate and often retract away from the injury site. Axons spared from the initial injury may later undergo secondary axonal degeneration. Lack of growth cone formation, regeneration, and loss of additional myelinated axonal projections within the spinal cord greatly limits neurological recovery following injury. To assess how central myelinated axons of the spinal cord respond to injury, we developed an ex vivo living spinal cord model utilizing transgenic mice that express yellow fluorescent protein in axons and a focal and highly reproducible laser-induced spinal cord injury to document the fate of axons and myelin (lipophilic fluorescent dye Nile Red) over time using two-photon excitation time-lapse microscopy. Dynamic processes such as acute axonal injury, axonal retraction, and myelin degeneration are best studied in real-time. However, the non-focal nature of contusion-based injuries and movement artifacts encountered during in vivo spinal cord imaging make differentiating primary and secondary axonal injury responses using high resolution microscopy challenging. The ex vivo spinal cord model described here mimics several aspects of clinically relevant contusion/compression-induced axonal pathologies including axonal swelling, spheroid formation, axonal transection, and peri-axonal swelling providing a useful model to study these dynamic processes in real-time. Major advantages of this model are excellent spatiotemporal resolution that allows differentiation between the primary insult that directly injures axons and secondary injury mechanisms; controlled infusion of reagents directly to the perfusate bathing the cord; precise alterations of the environmental milieu (e.g., calcium, sodium ions, known contributors to axonal injury, but near impossible to manipulate in vivo); and murine models also offer an advantage as they provide an opportunity to visualize and manipulate genetically identified cell populations and subcellular structures. Here, we describe how to isolate and image the living spinal cord from mice to capture dynamics of acute axonal injury.

An Ex Vivo Laser-induced Spinal Cord Injury Model to Assess Mechanisms of Axonal Degeneration in Real-time

We present a protocol utilizing two-photon excitation time-lapse microscopy to simultaneously visualize the dynamics of axon and myelin injuries in real time. This proposed protocol permits studies of both intrinsic and extrinsic factors which can influence central myelinated axon fate after injury and contribute to permanent clinical disability.

一个<em&gt;离体</em&gt;激光诱导脊髓损伤模型，以评估轴索变性的机制在实时

Injured CNS axons fail to regenerate and often retract away from the injury site. Axons spared from the initial injury may later undergo secondary axonal ...

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in adult mammalian animals remains elusive. In this study, we describe an experimental procedure for measuring calcium dynamics through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy. This method enables recordings and quantifications of neural activity in bilateral mouse brain regions one at a time in the same experiment for a prolonged period in vivo. Key aspects of this method, which can be completed within an hour, include minimally invasive surgery procedures for creating dual optical windows, and the use of 2P imaging. Although we only demonstrate the technique in the S1 area, the method can be applied to other regions of the living brain facilitating the elucidation of structural and functional complexities of brain neural networks.

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo.

胸腺1-gmcmp6 转基因小鼠在原代体血源皮质中的成像神经活性

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...

A Neurosphere Assay to Evaluate Endogenous Neural Stem Cell Activation in a Mouse Model of Minimal Spinal Cord Injury

Neural stem cells (NSCs) in the adult mammalian spinal cord are a relatively mitotically quiescent population of periventricular cells that can be studied in vitro using the neurosphere assay. This colony-forming assay is a powerful tool to study the response of NSCs to exogenous factors in a dish; however, this can also be used to study the effect of in vivo manipulations with the proper understanding of the strengths and limitations of the assay. One manipulation of the clinical interest is the effect of injury on endogenous NSC activation. Current models of spinal cord injury provide a challenge to study this as the severity of common contusion, compression, and transection models cause the destruction of the NSC niche at the site of the injury where the stem cells reside. Here, we describe a minimal injury model that causes localized damage at the superficial dorsolateral surface of the lower thoracic level (T7/8) of the adult mouse spinal cord. This injury model spares the central canal at the level of injury and permits analysis of the NSCs that reside at the level of the lesion at various time points following injury. Here, we show how the neurosphere assay can be utilized to study the activation of the two distinct, lineally-related, populations of NSCs that reside in the spinal cord periventricular region - primitive and definitive NSCs (pNSCs and dNSCs, respectively). We demonstrate how to isolate and culture these NSCs from the periventricular region at the level of injury and the white matter injury site. Our post-surgical spinal cord dissections show increased numbers of pNSC and dNSC-derived neurospheres from the periventricular region of injured cords compared to controls, speaking to their activation via injury. Furthermore, following injury, dNSC-derived neurospheres can be isolated from the injury site &#8212; demonstrating the ability of NSCs to migrate from their periventricular niche to sites of injury.

Here, we demonstrate the performance of a minimal spinal cord injury model in an adult mouse that spares the central canal niche housing endogenous neural stem cells (NSCs). We show how the neurosphere assay can be used to quantify activation and migration of definitive and primitive NSCs following injury.

小鼠脊髓损伤模型中内源性神经干细胞活化的干细胞检测

Neural stem cells (NSCs) in the adult mammalian spinal cord are a relatively mitotically quiescent population of periventricular cells that can be studied ...

Laminectomy and Spinal Cord Window Implantation in the Mouse

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated digital vaporizer is utilized to achieve a stable plane of anesthesia at a low-flow rate of isoflurane. A single vertebral spine is removed, and a commercially available cover-glass is overlaid on a thin agarose bed. A 3D-printed plastic backplate is then affixed to the adjacent vertebral spines using tissue adhesive and dental cement. A stabilization platform is used to reduce motion artifact from respiration and heartbeat. This rapid and clamp-free method is well-suited for acute multi-photon fluorescence microscopy. Representative data are included for an application of this technique to two-photon microscopy of the spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5 &#8212; a tight junction protein.

This protocol describes implantation of a glass window onto the spinal cord of a mouse to facilitate visualization by intravital microscopy.

拉米内切手术和脊髓窗口植入小鼠

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated ...

Activity-based Training on a Treadmill with Spinal Cord Injured Wistar Rats

Spinal cord injury (SCI) results in lasting deficits that include both mobility and a multitude of autonomic-related dysfunctions. Locomotor training (LT) on a treadmill is widely used as a rehabilitation tool in the SCI population with many benefits and improvements to daily life. We utilize this method of activity-based task-specific training (ABT) in rodents after SCI to both elucidate the mechanisms behind such improvements and to enhance and improve upon existing clinical rehabilitation protocols. Our current goal is to determine the mechanisms underlying ABT-induced improvements in urinary, bowel, and sexual function in SCI rats after a moderate to severe level of contusion. After securing each individual animal in a custom-made adjustable vest, they are secured to a versatile body weight support mechanism, lowered to a modified three-lane treadmill and assisted in step-training for 58 minutes, once a day for 10 weeks. This setup allows for the training of both quadrupedal and forelimb-only animals, alongside two different non-trained groups. Quadrupedal-trained animals with body weight support are aided by a technician present to assist in stepping with proper hind limb placement as necessary, while forelimb-only trained animals are raised at the caudal end to ensure no hind limb contact with the treadmill and no weight-bearing. One non-trained SCI group of animals is placed in a harness and rests next to the treadmill, while the other control SCI group remains in its home cage in the training room nearby. This paradigm allows for the training of multiple SCI animals at once, thus making it more time-efficient in addition to ensuring that our pre-clinical animal model mimics the clinical representation as close as possible, particularly with respect to the body weight support with manual assistance.

This protocol demonstrates our model of activity-based locomotor treadmill training for rats with spinal cord injury (SCI). Included is both quadrupedal and forelimb-only groups, in addition to two distinct types of non-trained control groups. Investigators are able to assess training effects on SCI rats using this protocol.

脊髓损伤 wistar 大鼠跑步机的作业训练

Spinal cord injury (SCI) results in lasting deficits that include both mobility and a multitude of autonomic-related dysfunctions. Locomotor training (LT) ...

Thoracic Spinal Cord Hemisection Surgery and Open-Field Locomotor Assessment in the Rat

Spinal cord injury (SCI) causes disturbances in motor, sensory, and autonomic function below the level of the lesion. Experimental animal models are valuable tools to understand the neural mechanisms involved in locomotor recovery after SCI and to design therapies for clinical populations. There are several experimental SCI models including contusion, compression, and transection injuries that are used in a wide variety of species. A hemisection involves the unilateral transection of the spinal cord and disrupts all ascending and descending tracts on one side only. Spinal hemisection produces a highly selective and reproducible injury in comparison to contusion or compression techniques that is useful for investigating neural plasticity in spared and damaged pathways associated with functional recovery. We present a detailed step-by-step protocol for performing a thoracic hemisection at the T8 vertebral level in the rat that results in an initial paralysis of the hindlimb on the side of the lesion with graded spontaneous recovery of locomotor function over several weeks. We also provide a locomotor scoring protocol to assess functional recovery in the open-field. The locomotor assessment provides a linear recovery profile and can be performed both early and repeatedly after injury in order to accurately screen animals for appropriate time points in which to conduct more specialized behavioral testing. The hemisection technique presented can be readily adapted to other transection models and species, and the locomotor assessment can be used in a variety of SCI and other injury models to score locomotor function.

The rat thoracic spinal hemisection is a valuable and reproducible model of unilateral spinal cord injury to investigate the neural mechanisms of locomotor recovery and treatment efficacy. This article includes a detailed step-by-step guide to perform the hemisection procedure and to assess locomotor performance in an open-field arena.

胸脊髓截断手术和大鼠开放场运动评估

Spinal cord injury (SCI) causes disturbances in motor, sensory, and autonomic function below the level of the lesion. Experimental animal models are ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to intracranial delivery, intra-arterial administration of NSCs is less invasive and produces a more diffuse distribution of NSCs within the brain parenchyma. Further, intra-arterial delivery allows the first-pass effect in the brain circulation, lessening the potential for trapping of cells in peripheral organs, such as liver and spleen, a complication associated with peripheral injections. Here, we detail the methodology, in both mice and rats, for delivery of NSCs through the common carotid artery (mouse) or external carotid artery (rat) to the ipsilateral hemisphere after an ischemic stroke. Using GFP-labeled NSCs, we illustrate the widespread distribution achieved throughout the rodent ipsilateral hemisphere at 1 d, 1 week and 4 weeks after postischemic delivery, with a higher density in or near the ischemic injury site. In addition to long-term survival, we show evidence of differentiation of GFP-labeled cells at 4 weeks. The intra-arterial delivery approach described here for NSCs can also be used for administration of therapeutic compounds, and thus has broad applicability to varied CNS injury and disease models across multiple species.

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

神经干细胞向大鼠和小鼠大脑的动脉内传递：对脑缺血的应用

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...

Establishing a Mouse Contusion Spinal Cord Injury Model Based on a Minimally Invasive Technique

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, thereby improving the reproducibility of the experiments.
These methods need two requirements to be fulfilled: clarity of the surgical anatomical pathway and simplicity and convenience of the laboratory device. Crucially for the operator, a clear anatomical pathway provides minimally invasive exposure, which avoids additional damage to the experimental animal during the surgical procedures and allows the animal to maintain a consistent and stable anatomical morphology during the experiment.
In this study, the use of a novel integrated platform called the SCI coaxial platform for spinal cord injury in small animals to expose the T9 level spinal cord in a minimally invasive way and stabilize and immobilize the vertebra of mice using a vertebral stabilizer is researched, and, finally, a coaxial gravity impactor is used to contuse the spinal cord of mice to approach different degrees of T9 spinal cord injury. Finally, histological results are provided as a reference for the readers.

Minimally invasive techniques and a simple laboratory device improve the reproducibility of the spinal cord injury model by reducing operative damage to the experimental animals and allowing anatomical morphology maintenance. The method is worthwhile because the reliable results and reproducible procedure facilitate investigations of the mechanisms of disease reparation.

基于微创技术的小鼠挫伤脊髓损伤模型的建立

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, ...